Investigation of the species-dependent in vitro metabolism of BAL30630 by stable isotope labeling and isotope exchange experiments analyzed by capillary liquid chromatography coupled to mass spectrometry
| Autor: | Anne Schmitt-Hoffmann, Dominique Klauer, Mathias Wind, Patrick Roussel, Helge Grunwald, Michael Schleimer, Pascal Fullhardt, Jochen Spickermann, Klaus Illig, Claude Nuoffer, Klaus Gebhardt |
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| Rok vydání: | 2009 |
| Předmět: |
Antifungal Agents
Metabolite Guinea Pigs 1-Propanol Tandem mass spectrometry Mass spectrometry Biochemistry Mass Spectrometry Analytical Chemistry Mice chemistry.chemical_compound Dogs Liquid chromatography–mass spectrometry Animals Humans Chromatography Isotope Chemistry Stable isotope ratio Organic Chemistry Haplorhini General Medicine Deuterium Rats Isotope Labeling Hepatocytes Microsomes Liver Microsome Hydrogen–deuterium exchange Rabbits Chromatography Liquid |
| Zdroj: | Journal of Chromatography A. 1216:3946-3953 |
| ISSN: | 0021-9673 |
| Popis: | The in vitro metabolic profile of BAL30630, an antifungal piperazine propanol derivative, which inhibits the 1,3-beta- d -glucansynthase, was investigated by incubation with microsomes of several species and with rat hepatocytes. For the spotting of the metabolites, mixtures of BAL30630 with a stable isotope (deuterium) labeled analogue were incubated. The metabolic pattern comprises several oxidized metabolites. Based on isotope exchange experiments, their structures could be assigned to epoxide- and hydroxylated metabolites. In hepatocyte incubations, several glucuronides formed from these oxidized metabolites could be observed. From the analysis of the metabolic pattern in microsomes, products of carbamate hydrolysis were characterized. This hydrolysis was highly species dependent. In activated incubations and in rat hepatocytes, those metabolites were further oxidized. In incubations without NADPH activation, the resulting hydrolytic metabolites could be enriched without the subsequent oxidation. Final structural elucidation of the metabolites was performed using accurate mass determination and isotope exchange experiments, in which incubations were analyzed by deuterium exchange and capillary HPLC–QTof-MS and MS/MS. The use of non-radioactive, stabile isotope labeled drug analogues in combination with isotope exchange studies was essential in particular for a defined assignment of the functional groups in the structures of the investigated metabolites. |
| Databáze: | OpenAIRE |
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